Electrophoresis Using Cleangel And Buffer Strips - Pharmacia Biotech Multiphor II User Manual

5.2 Electro-
phoresis
using
CleanGel and
buffer strips
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Time schedule for silver staining
Step No. Solution
1 Fixing solution C
2 Incubation solution D
3 Distilled water
4 Silver solution E
5 Developing solution F
6 Stop solution G
7 Distilled water
8 Preserving solution L
*Short development times will give a lightly stained gel. Long development times will give a
dark gel.
To preserve the gel, soak a cellophane sheet in preserving solution (L) and
lay it on the gel surface. Remove any air bubbles and wrap the excess
around the glass plate. An additional glass plate may be placed underneath
during drying to stop the cellophane from shifting or wrinkling. Leave the
gel at room temperature until it is completely dry.
This section gives a brief description of the running procedure for native
PAGE and SDS PAGE using CleanGel and CleanGel buffer strips.
CleanGel is a washed and dried polyacrylamide gel. Depending on the
application, the gels are rehydrated in different buffers to a gel thickness of
0.5 mm. The gels are available with various numbers and sizes of sample
wells. There are various application kits with prepared buffers. For more
information, see Ordering information and the information supplied with
the CleanGel.
Preparing the experiment
Connect Multiphor II to MultiTemp II thermostatic circulator. Switch on
MultiTemp II 20 minutes before starting the experiment and set the
temperature to 15 °C.
Note: Always wear clean gloves when working with polyacrylamide gels
and buffer strips, particularly when using sensitive staining methods.
5. Operation
Time (min)
30
30
3x5
40
5-15 *
10
3x5
20
25